Streptavidin-Enzyme Linked Aggregates for the One-Step Assembly and Purification of Enzyme Cascades

Mallin, Hendrik and Ward, Thomas R.. (2018) Streptavidin-Enzyme Linked Aggregates for the One-Step Assembly and Purification of Enzyme Cascades. ChemCatChem, 10 (13). pp. 2810-2816.

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Official URL: https://edoc.unibas.ch/70536/

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Herein, we report enzyme aggregates assembled around covalently cross-linked streptavidin tetramers. The streptavidin oligomeric matrix (Sav(Matrix)) is produced by using SpyTag/SpyCatch technology and binds tightly to fusion proteins bearing a streptavidin-binding peptide (SBP). Fusing the SBPs to different enzymes leads to precipitation of the streptavidin-enzyme aggregates upon mixing the complementary components. This straightforward strategy can be applied to crude cell-free extracts, allowing the one-step assembly and purification of catalytically active aggregates. Enzyme cascade assemblies can be produced upon adding different SBP-fused enzymes to the Sav(Matrix). The reaction rate for lactate dehydrogenase (LDH) is improved tenfold (compared with the soluble enzyme) upon precipitation with the Sav(Matrix) from crude cell-free extracts. Additionally, the kinetic parameters are improved. A cascade combining a transaminase with LDH for the synthesis of enantiopure amines from prochiral ketones displays nearly threefold rate enhancement for the synthesis of (R)-alpha-methylbenzylamine compared with the free enzymes in solution.
Faculties and Departments:05 Faculty of Science > Departement Chemie > Chemie > Bioanorganische Chemie (Ward)
UniBasel Contributors:Ward, Thomas R. R. and Mallin, Hendrik
Item Type:Article, refereed
Article Subtype:Research Article
Note:Publication type according to Uni Basel Research Database: Journal article
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Last Modified:08 Feb 2020 15:09
Deposited On:22 May 2019 08:50

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